술패랭이꽃(Dianthus superbus)의 엽육원형질체로부터 식물체 재분화

Plant Regeneration from Mesophyll Protoplasts of Dianthus superbus

술패랭이꽃의 엽육원형질체를 2.0mg/L NAA, 0.5 mg/L BAP 및 9%의 mannitol이 포함된 MSPI 9M 액체배지에서 배양하였다. 27$^{\circ}C$, 암조건에서 배양 3-4주일 후 활발한 분열과정을 거쳐 원형질체로부터 colony가 형성되었으며 이들colony는 연속광 (21.5 $\mu$Eㆍ $m^{-2}$ ㆍse $c^{-1}$)하에서 배양 2주일 후 직경 약 3 mm의 녹색 microcallus로 생장하였다. 녹색 microcallus는 2.0 mg/L 2,4-D가 첨가된 고체배지에서 배양30일 후 배발생 캘러스를 형성하였으며 이들 배발생 캘러스는 0.1 mg/L 2,4-D, 2.0 mg/L kinetin 및 2.0 g/L casein hydrolysate가 포함된 $N_{6-}$2 배지에서 캘러스당 10-15개의 multiple shoot의 분화가 이루어졌다. 재분화된 shoot는 2.0mg/L NAA가 포함된 MS 배지에서 뿌리가 유도되었으며 이들을 pot로 이식하여 재분화 개체를 획득할 수 있었다.

Leaf mesophyll protoplasts of Dianthus superbus were cultured in MSP1 liquid medium supplemented with 0.5 mg/L BAP, 2.0 mg/L NAA and 9% mannitol. Protoplast-derived colonies were formed after 3 to 4 weeks of culture in the dark at 27$^{\circ}C$. These colonies were kept under continuous illumination (21.5 $\mu$E. m-2 sec-1) for 2 weeks and finally most of the colonies became green microcalli, about 3 mm in diameter. When green microcalli were transferred to MS solidified medium with 2.0 mg/L 2,4-D, they formed embryogenic calli after 4 week of culture. These calli were then transferred onto $N_{6}$ medium containing 0.1mg/L 2,4-D, 0.1 mg/L NAA, 2.0 mg/L kinetin and 2.0 g/L casein hydrolysate and cultured under illumination. After 5 weeks of culture the calli gave rise to multiple shoots of 10 to 15 per callus. Upon transfer onto MS medium containing 2.0 mg/L NAA, they were noted. The regenerates were successfully transplanted into potting soil.